GenCRISPR™ ゲノム編集関連製品 » GenCRISPR™ ヌクレアーゼ-CRISPR遺伝子編集 » GenCRISPR™ Cas12a (Cpf1) Nuclease
GenCRISPR™ Cas12a (Cpf1) Nuclease

A 20 μl reaction in 1xCas12a Nuclease Reaction Buffer containing 60 ng linearized plasmid, 10 ng crRNA, and 100 ng GenCRISPR™ Cas12a (Cpf1) Nuclease for 30 mins at 37°C results in a digestion efficiency of linearized plasmid higher than 90%, as determined by agarose gel electrophoresis.

GenCRISPR™ Cas12a (Cpf1) Nuclease

The clustered regularly interspaced short palindromic repeats, known as CRISPR systems are adaptive immune mechanisms commonly present in archaea and bacteria. The CRISPR systems enable the host to specifically target and cleave foreign nucleic acids thus targeting infectiousviruses and plasmids. Recently, a type V CRISPR system has been identified in several bacteria, the Cpf1 CRISPR from Prevotella and Francisella 1. In contrast to Cas9 systems, CRISPR/Cpf1 systems are smaller in size, do not require an additional trans-activating crRNA (tracrRNA), and allow for targeting of additional genomic regions by cleaveing the target DNA proceeded by a short T-rich protospacer-adjacent motif (PAM). On the other hand, the Cas9 system requires a G-rich PAM following the target DNA. Furthermore, Cas12a/Cpf1 introduces a staggered DNA double stranded break with a 4 or 5-nt 5’ overhang. Recombinant Acidaminococcus sp. BV3L6 Cas12a (cpf1) nuclease is expressed in E. coli and purified. The nuclease contains nuclear localization sequence (NLS) at the C-terminus and 6× His-tag at the C-terminus.
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Description

The_x000D_ clustered regularly interspaced short palindromic repeats, known as CRISPR  systems are adaptive immune mechanisms commonly_x000D_ present in archaea and bacteria. The CRISPR systems enable the host to specifically_x000D_ target and cleave foreign nucleic acids thus targeting infectiousviruses and_x000D_ plasmids. Recently, a type V CRISPR system has been identified in several_x000D_ bacteria, the Cpf1 CRISPR from Prevotella and Francisella 1. In contrast to_x000D_ Cas9 systems, CRISPR/Cpf1 systems are smaller in size, do not require an_x000D_ additional trans-activating crRNA (tracrRNA), and allow for targeting of_x000D_ additional genomic regions by cleaveing the target DNA proceeded by a short_x000D_ T-rich protospacer-adjacent motif (PAM). On the other hand, the Cas9 system requires_x000D_ a G-rich PAM following the target DNA. Furthermore, Cas12a/Cpf1 introduces a_x000D_ staggered DNA double stranded break with a 4 or 5-nt 5’ overhang. Recombinant_x000D_ Acidaminococcus sp. BV3L6 Cas12a (cpf1) nuclease is expressed in E. coli and_x000D_ purified. The nuclease contains nuclear localization sequence (NLS) at the_x000D_ C-terminus and 6× His-tag at the C-terminus.
Note

For laboratory research use only. Direct human use, including taking orally and injection and clinical use are forbidden.

Applications
crRNA -dependent double-stranded DNA cleavage

Properties
Expression System Recombinant Cas12a with an C-terminal NLS expressed by E.coli
Species Acidaminococcus sp. (strain BV3L6)
Tag C-terminal 6× His Tag
Apparent Molecular Weight ~150 kDa, on SDS-PAGE under reducing conditions
Concentration 4 mg/ml
Active temperature This Cas12a is active at 37℃.
Formulation Supplied as a solution of 10 mM Tris, pH7.4, 300 mM NaCl, 0.5 mM DTT, and 50% glycerol. 
Storage & Stability This product remains stable up to 18 months at -20℃. Avoid repeated freeze-thaw cycles.
Accession# U2UMQ6
Key Features High knockout efficiencies: Consistent high performance in in-vitro plasmid cleavage test.
Time-saving: no need for transcription and translation. 
DNA-free: no external DNA added to system.
Quality Control Specifications
AssaySpecifications
AppearanceClear, colorless liquid
Purity by SDS-PAGE≥ 90%
Concentration by A2804(+ 10%) mg/ml
Bioactivity ( in vitro)≥ 90%
Residual DNaseUndetectable
Residual RNaseUndetectable
Endotoxin Level≤ 0.1 EU/μg of the protein by gel clotting method

Examples
  • GenCRISPR™ Cas12a (Cpf1) Nuclease
    • GenCRISPR™ Cas12a (Cpf1) Nuclease

    A 20 μl reaction in 1xCas12a Nuclease Reaction Buffer containing 60 ng linearized plasmid, 10 ng crRNA, and 100 ng GenCRISPR™ Cas12a (Cpf1) Nuclease for 30 mins at 37°C results in a digestion efficiency of linearized plasmid higher than 90%, as determined by agarose gel electrophoresis.